PROJECT SUMMARY Chronic pain occurs frequently after burn injury and spinal cord injury (SCI). Currently available treatments are often ineffective or only partially effective. Relatively little is known about the mechanisms responsible for the onset and persistence of pain following burn injury, despite chronic pain being a frequent complaint of burn- injured and SCI patients. Our goal is to identify and characterize cellular and molecular mechanisms that contribute to pain following burn and spinal cord injuries, with the objective of delineating specific targets for more effective pain management. Our recent progress includes development of a burn injury model in rats that produces long-lasting mechanical allodynia associated with hyperexcitability of spinal cord dorsal horn (DH) neurons, and demonstration that pain and DH hyperexcitability occur concomitant with activation of DH microglia, with the spread of pain paralleled by spreading microglial activation in this model. In addition, we have shown that acute early inhibition of microglial activation attenuates burn-induced mechanical allodynia and DH neuronal hyperexcitability. We have also demonstrated that the maintenance of below-level pain following SCI is associated with activation of microglia, and that some microglial functions are regulated by Na channels and can be attenuated with Na channel blockade. Recently, we have also shown that activated polymorphonuclear neutrophils significantly increase excitability of DRG neurons, as manifested by lowered threshold and increased firing frequency. Our preliminary data indicate that macrophages infiltrate DRG following burn injury and SCI, at a time when pain behavior is evident. We have also shown hyperexcitability in DH neurons in vivo in conjunction with activation of DH microglia following burn injury and SCI. We now plan to build upon our progress, via the following specific aims. 1. Elucidate the molecular changes in DRG and DH neurons, and glia, following burn injury, and examine novel pharmacotherapeutic approaches to pain following burn injury. 2. Investigate the effect of Na channel blockade on the activity of microglia following SCI, and determine whether channel blockade can reduce microglial activation and attenuate pain behavior. 3. Examine if infiltration of macrophages into at- and/or below-level DRG is associated with the development and/or persistence of neuropathic pain following SCI. 4. Determine whether Na channel blockade attenuates macrophage infiltration into DRG following SCI and whether neutralizing TNF-a reduce macrophage infiltration into DRG following SCI. 5. Investigate the effects of macrophages and microglia on the excitability of DRG and DH neurons, and determine whether DRG neuron hyperexcitability following activation of these cells can be prevented via neutralization of cytokines which are expressed by immune cells. PUBLIC HEALTH RELEVANCE: There is a very substantial need, within the VA and within the U.S. population in general, for more effective treatments for pain associated with burn injury and spinal cord injuries. Relatively little is known of the mechanisms responsible for the onset and persistence of pain following burn injury despite chronic pain being the most frequent complaint of burn-injured patients. Chronic pain and dysethesiae that develop following burn injury, nerve and spinal cord injuries are often resistant to conventional therapeutic approaches. These forms of pain represent significant therapeutic challenges, often being unresponsive or only partially responsive, to existing therapies. Our goal is to identify and characterize cellular and molecular mechanisms that contribute to pain following burn and spinal cord injuries, with the objective of delineating specific targets for more effective pain management.